Nucleic acid separation on the QIAxcel Advanced System, using the QX DNA High Resolution Kit takes less than 10% of the time of conventional high-resolution agarose.

Streamlined RNA analysis using the QIAxcel Advanced System.

Total RNA purified from Schizosaccharomyces pombe. Results presented as [A] a gel image and [B] a superimposed electropherogram view. Reliable quality control of unlabeled cRNA generated from yeast total RNA. Results presented as [C] a gel image and [D] a superimposed electropherogram view.

RIS indicates RNA integrity.

Total RNA was extracted from rat liver samples using the RNeasy Plus Mini Kit and subjected to decomposing conditions. Samples of increasing levels of degradation were analyzed on the QIAxcel Advanced and the Agilent 2100 Bioanalyzer platform. The results are depicted in the gel view pictures for the QIAxcel Advanced [A] and the Agilent Bioanalyzer [B]. The electropherogram of selected samples analyzed on the QIAxcel Advanced is shown in [C]. The RIS (QIAxcel Advanced) and RIN (Agilent Bioanalyzer) numbers are listed in the table.

Accurate, high-throughput genotyping of bacteria.

Samples were analyzed on either [A] the QIAxcel Advanced System at the original concentration or [B] by agarose gel electrophoresis with five-fold concentration. Samples 3 [C] and 5 [D] were analyzed by agarose gel electrophoresis at the original concentration (O), with five-fold concentration (C), and at the original concentration on the QIAxcel Advanced (Q). The arrows represent bands originally scored as negative. Data kindly provided by Mutschall and coworkers, Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Lethbridge, Alberta, Canada.

Nucleic acid molecules are size separated by applying a current to a gel-filled capillary, and detected as they migrate toward the positively charged terminus. The signal data pass through a photomultiplier and are converted to an electropherogram and gel image by the QIAxcel ScreenGel Software.

Streamlined workflow

The QIAxcel Advanced system not only speeds up DNA and RNA analysis by eliminating slab-gel analysis, but also contributes to streamlining the entire sample purification and analysis workflow. Used in combination with proven QIAGEN end-point PCR kits, the QIAxcel Advanced provides an all-in-one, pretested solution for reliable analysis of PCR fragments, ensuring reproducibility and significant time and cost savings.

One instrument, many applications

The QIAxcel Advanced system is a versatile solution for electrophoresis and offers a broad range of applications. Preprogrammed methods, in combination with the corresponding gel cartridges, allow separation and analysis of a variety of nucleic acids — including single or multiple PCR fragments, DNA digested with restriction endonucleases, total RNA, and cRNA. Unlike other commercially available devices that provide processing of only up to 12 samples at time, the QIAxcel Advanced system allows processing of all 96 samples without manual intervention.

RNA analysis — objective quality values with the RNA Integrity Score

The new QIAGEN RNA Integrity Score (RIS) provides an objective quality measurement for eukaryotic RNA samples and allows easy interpretation of sample integrity. The RIS is a value from 1 to 10 where a value of 10 indicates completely intact RNA (see figure RIS indicates RNA integrity). The highly reproducible RIS eliminates the need for human interpretation and enables the implementation of rigorous quality control at appropriate breakpoints across workflows. This allows researchers to eliminate samples that do not meet the quality criteria required for downstream processes, reducing the risk of needing to repeat experiments and saving time and costs.

Such careful monitoring of the initial RNA sample, as well as products generated throughout the entire procedure, is crucial in microarray analysis, since RNA degradation strongly influences the predictive power obtained from the microarray data. The QIAxcel Advanced System is highly suitable for analyzing the integrity of total RNA and size distribution of fragmented or intact cRNA (see figure Streamlined RNA analysis using the QIAxcel system), and the throughput capacity makes it very convenient for laboratories employing 96-well RNA purification technologies that need a fast solution for quality control.

Principle

The QIAxcel Advanced instrument includes an array of light-emitting diodes and micro-optical collectors that latch to capillaries within QIAxcel gel cartridges. Fragments that are migrating through a gel matrix within the capillary pass excitation and detection spots and the signal is transmitted through a photomultiplier tube to the QIAxcel ScreenGel software for data interpretation (see figure Sample separation process).

The high detection sensitivity provided by the QIAxcel Advanced instrument enables robust results even with low concentrations of nucleic acid. With a resolution of 3–5 bp for fragments smaller than 0.5 kb, the QIAxcel Advanced system ensures greater accuracy than slab-gel methods, as well as greater confidence in data interpretation. Sample consumption is less than 0.1 μl per analysis, saving precious samples for further downstream analysis. Hands-free sample loading and self-contained components minimize exposure to hazardous chemicals, such as ethidium bromide (see figure Ready-to-run gel cartridges).

A range of QIAxcel Kits is available for a variety of applications, meeting different resolution and speed requirements (see table below).

QIAxcel Kits

QIAxcel Kit

Analyte

Size range

100 bp – 500 bp*

500 bp – 1 kb*

1 kb – 5 kb*

5 kb – 10 kb*

Run time/
12 samples†

QIAxcel DNA High Resolution Kit

DNA

15 bp–10 kb

3–5 bp

50 bp

200–500 bp

1–1.5 kb

7–20 min

QIAxcel DNA Screening Kit

DNA

15 bp–5 kb

20–50 bp

50–100 bp

500 bp

–

5 min

QIAxcel DNA Fast Analysis Kit

DNA

15 bp–3 kb

50–100 bp

100–250 bp

250 bp – 1 kb‡

–

3–5 min

QIAxcel RNA QC Kit v2.0

RNA

15 b–6 kb

–

–

–

–

10 min

* Best resolution. † Run time depends on method used. ‡ Best resolution between 1–3 kb.

Procedure

Just a few, simple steps are required to operate the QIAxcel Advanced system for analysis of DNA and RNA samples: load the gel cartridge, fill and load the buffer tray, load the samples in 96-well plates or in PCR tubes or strips, select the process profile to be used — and go! Tedious gel preparation and extensive user training are eliminated, streamlining the workflow and facilitating integration of the system into the daily lab routine. Within minutes of starting a run, the first results appear in real time on the computer screen (see figure Reduced run times).

Applications

The QIAxcel Advanced System offers a broad range of optimized applications for DNA and RNA analysis and is highly suited for research in academic as well as pharmaceutical, biotechnology, and biomedical laboratories. Preprogrammed methods, in combination with the corresponding gel cartridges, allow separation and analysis of a variety of nucleic acids — including single or multiple PCR fragments, DNA digested with restriction endonucleases, synthesized oligonucleotides, total RNA, and cRNA.

The QIAxcel Advanced System has been used for applications such as:

Bacterial genotyping

PCR fragment analysis

Pre-sequencing testing

Mutation detection

Detection of food allergens

Quality control of genomic DNA

Total RNA and cRNA QC for RT-PCR and microarray analysis

Software

QIAxcel ScreenGel software provides a new level of convenience

QIAxcel ScreenGel software, specifically developed for use with the QIAxcel Advanced system, is a powerful and user-friendly tool for data collection and analysis. Interactive tools simplify analysis, facilitate rapid data interpretation, and provide flexibility with data and results displayed in both electropherogram and gel image formats. Results can be viewed individually or displayed as overlay views for sample and data comparison. All-in-one analysis for multiple data sets simplifies evaluation. A unique algorithm calculates and generates a tabular display of a variety of peak properties, including number of peaks as well as the size, height, width, and area of each peak. Comprehensive data reports can be easily generated and saved or exported to meet individual documentation needs. Complete process profiles for standardized sample processing — from running samples to data analysis, generating reports and exporting data — minimize the need for additional training for users (see figure Simplified and standardized data collection and analysis).

Easy setup and start

The Process Wizard simplifies experiment startup (see figure Easy-to-use software features). This convenient feature enables guided setup and allows run parameters to be defined and DNA size markers to be selected. Reagent lot number information can be included. Samples can be selected with a single click of the mouse and run checks can also be easily performed.

New RNA Integrity Score

The updated QIAxcel ScreenGel software now features the QIAGEN RNA Integrity Score (RIS), which enables objective quality measurement of analyzed samples and allows easy interpretation of sample integrity. The RIS is a value from 1 to 10 where a value of 10 indicates completely intact RNA (see figure RIS indicates RNA integrity). The highly reproducible RIS eliminates the need for human interpretation and enables the implementation of rigorous quality control at appropriate breakpoints across workflows. This allows researchers to eliminate samples that do not meet the quality criteria required for downstream processes, reducing the risk of needing to repeat experiments and saving time and costs.

Support of 21 CFR Part 11 compliance

QIAxcel ScreenGel software includes specific features that support the technical requirements of 21 CFR Part 11 regulations. Security features such as the following are available:

Different user profiles are available (Routine, Basic, Advanced, Admin) and are password protected (user login is required) for increased security. A simplified interface minimizes the need for user training and makes the software especially attractive for inexperienced users.